This manuscript is contextually identical with the following published paper: Kuszegi, G., Siller, I., Dima, B., Takács, K., Merényi, Zs., Varga, T., Turcsányi, G., Bidló, A., Ódor, P. 2015. Drivers of macrofungal species composition in temperate forests, West Hungary: functional groups compared. Fungal Ecology 17: 69-83. DOI: 10.1016/j.funeco.2015.05.009 The original published pdf available in this website: http://authors.elsevier.com/sd/article/S0378112713004295 Title: Drivers of macrofungal species composition in temperate forests, West Hungary: functional groups compared Authors: Gergely Kutszegi1,*, Irén Siller2, Bálint Dima3, 6, Katalin Takács3, Zsolt Merényi4, Torda Varga4, Gábor Turcsányi3, András Bidló5, Péter Ódor1 1MTA Centre for Ecological Research, Institute of Ecology and Botany, Alkotmány út 2–4, H-2163 Vácrátót, Hungary, [email protected], [email protected]. 2Department of Botany, Institute of Biology, Szent István University, P.O. Box 2, H-1400 Budapest, Hungary, [email protected]. 3Department of Nature Conservation and Landscape Ecology, Institute of Environmental and Landscape Management, Szent István University, Páter Károly út 1, H-2100 Gödöllő, Hungary, [email protected], [email protected], [email protected]. 4Department of Plant Physiology and Molecular Plant Biology, Eötvös Loránd University, 1 Pázmány Péter sétány 1/C, H-1117 Budapest, Hungary, [email protected], [email protected]. 5Department of Forest Site Diagnosis and Classification, University of West-Hungary, Ady út 5, H-9400 Sopron, Hungary, [email protected]. 6Department of Biosciences, University of Helsinki, P.O. Box 65, 00014 University of Helsinki, Helsinki, Finland, [email protected]. * corresponding author, Fax: +36 28 360110, email address: [email protected] (Gergely Kutszegi) Footnote to the title: Environmental drivers of macrofungal species composition 2 Abstract The most influential environmental drivers of macrofungal species composition were studied in managed, even-aged, mixed forests of Őrség National Park, Hungary. Functional groups of macrofungi were analyzed separately by non-metric multidimensional scaling and redundancy analysis exploring their relations to tree species composition, stand structure, soil/litter conditions, microclimate, landscape, and management history. Some evidence was given that macrofungi are related to drivers that are relatively easy to measure. It was found that wood- inhabiting fungal species composition is driven primarily by the species composition of living trees, while substrate properties and microclimate play minor roles. The terricolous saprotrophic community was determined principally by a litter pH gradient involving tree species composition and soil/litter properties. Microclimate had no concordant effect. No obvious underlying gradients were detected on ectomycorrhizal fungal species composition; however, tree size and litter pH had significant effects. For each group, no clear responses to landscape or management history were detected. Key words: biodiversity; ectomycorrhizal fungi; environmental variation; fungal community gradients; host specificity; soil properties; sporocarp sampling; terricolous saprotrophic fungi; wood-inhabiting fungi 3 Introduction Forest-dwelling macrofungal assemblages have been classified into three main functional groups: wood-inhabiting (including wood saprotrophs and necrotrophic parasites), ectomycorrhizal (EcM) and terricolous saprotrophic communities (Winterhoff, 1992). In a global perspective, an enormous volume of research has been reported on the responses of macrofungal community composition to environmental variation. It was revealed that wood- inhabiting fungi are driven principally by the amount and diameter (Heilmann-Clausen and Christensen, 2004; Sippola et al., 2005; Ódor et al., 2006; Lonsdale et al., 2008), decay stage (Heilmann-Clausen and Christensen, 2003b; Siller, 2004; Heilmann-Clausen et al., 2014), age (Heilmann-Clausen, 2001), species identity (Sippola et al., 2005; Küffer et al., 2008), complexity (Heilmann-Clausen and Christensen, 2003a), and spatio-temporal availability (Siitonen, 2001; Bässler et al., 2010; Halme et al., 2013) of dead wood. The microclimatic variation and pH within the wood (Boddy, 1992, 2001; Salerni et al., 2002) or the interactions with other organisms (van der Wal et al., 2013) had also significant effects. EcM community compositions were found to be structured strongly by the N content (Toljander et al., 2006; Cox et al., 2010; Suz et al., 2014), pH (Baar and ter Braak, 1996; Talbot et al., 2013) as well as temperature and moisture of soil (Claridge et al., 2000; Jones et al., 2003), species composition of host trees (Kernaghan et al., 2003; Smith and Read, 2008; Morris et al., 2009), season (over the course of even a month) (Courty et al., 2008), fungal dispersal limitation among host trees (Peay et al., 2010), and timing of colonization and interspecific competition on the root surface (Kennedy et al., 2009; Kennedy, 2010). In the same context, little is known about the determinants of terricolous saprotrophic communities, but the effects of litter quantity and pH (Tyler, 1991; Ferris et al., 2000; Talbot et al., 2013), P content of the soil (Reverchon et al., 2010), tree species composition (O’Hanlon and Harrington, 2012), and temperature (McMullan-Fisher et al., 2009) were documented to be highly important. 4 Many influential environmental drivers have been revealed, but are there drivers with consistent importance on macrofungal functional groups? When such drivers are sought, many difficulties are encountered. The relative importance of drivers varies across spatial scales (Claridge et al., 2000; Lilleskov and Parrent, 2007; Büntgen et al., 2012) and along environmental gradients, such as elevation (Gómez-Hernández et al., 2012; Sundqvist et al., 2013) and rainfall (Lindblad, 2001; Salerni et al., 2002). Also, the relative effects of drivers can be biased strongly by the edaphic heterogeneity of the studied habitats, and the actually limiting factors (resources or environmental conditions) in a habitat can have a disproportionately high influence on species composition (McMullan-Fisher, 2008). In addition, community level responses are difficult to reveal, since great species diversity is found within fungal communities in which each species have slightly different environmental requirements (Boddy et al., 2008). Based on the studies mentioned in the first paragraph, our knowledge of fungal community responses to environmental variation is biased by research history: (1) the majority of studies have been conducted in Northern or Western Europe or in North America, thus, large regions are still underrepresented; (2) the studies have rarely been focused on more than two functional groups (except e.g. Humphrey et al., 2000; Sato et al., 2012); (3) to obtain a clearer picture, many authors have used a limited pool of environmental factors and hence, several environmental impacts with probable significant effects remained unexplored on the sampling sites. Given this complexity and research gaps, the present study has been designed in even- aged, managed forests with a restricted number of habitat types to try to reduce the effects of edaphic heterogeneity. By including several variables suggested by the literature, other factors that characterize the landscape and management history were also examined. In accordance with the studies referenced in the first paragraph, it can be hypothesized 5 that (1) the effects of substrate properties, tree species composition, and microclimate have the strongest effects on macrofungal species composition at a stand scale, and (2) the relative influence of these factors differs among wood-inhabiting, EcM, and terricolous saprotrophic communities. The aims of this study are to find the most important environmental factors that best explain the macrofungal species composition of wood-inhabiting, EcM and terricolous saprotrophic communities, and provide information on the environmental requirements of fungal species. 6 Materials and methods Study area This study has been carried out in Őrség National Park (ŐNP), West Hungary (46° 51’–55’ North, 16° 06’–24’ East (Fig 1a). In the ŐNP, the precipitation ranges between 700 and 800 mm yearly. Between 1901 and 2000, the mean minimum and maximum temperatures in winter were respectively –7.4 and 6.0°C, while in summer 13.5 and 23.8°C (measured in a nearby town, Szombathely, Hungarian Meteorological Service, OMSZ). The landscape is divided into hills and wide valleys at the elevation range of 250–350 m above sea level. The bedrock consists of alluvial gravel and clay. Nutrient-poor brown forest soils with pseudogley or lessivage (planosols or luvisols) are the most frequent soil types (Halász, 2006; Dövényi, 2010). The pH of the soil is acidic; it tends to range from 4.0 to 4.8 with a mean of 4.3 (Juhász et al., 2011). Presently, forests cover 80% of the ŐNP region, which has an area of ca. 350 km2 (Dövényi, 2010). Stands are dominated by beech (Fagus sylvatica L.), sessile and pedunculate oak [Quercus petraea (Matuschka) Liebl. and Q. robur L.], hornbeam (Carpinus betulus L.), and Scots pine (Pinus sylvestris L.). Forests are sometimes monodominant, but more often form mixed stands with great compositional diversity. The most frequent non-dominant tree species are Betula pendula Roth, Picea abies (L.) Karst., Populus tremula L., Castanea